血清miR-187-3p对肝细胞癌诊断和预后判断临床价值研究

Clinical value of serum miR-187-3p on diagnosis and prognosis of hepatocarcinoma

目的 miRNA在血清中具有良好的稳定性和特异性,是近年来发现的一种在肿瘤诊断及预后判断中具有重要潜在应用价值的新型生物标志物。肝细胞癌患者血清中目前已发现多种具有诊断价值和预后相关的miRNAs。本研究旨在探索肝细胞癌患者血清miR-187-3p的相对含量与临床病理特征的相关性,分析血清miR-187-3p作为肝细胞癌生物标志物在诊断和预后中的价值。方法收集2012-01-01-2016-11-30于中国人民解放军成都军区总医院初诊为肝细胞癌的81例患者(患者未经任何治疗)血样,随机抽取同一时期年龄与性别基本一致的81名健康人血样作为对照。前期miRNA芯片筛选出Huh7肝癌细胞和Huh7肝癌干细胞中差异表达的miRNAs,并采用qRT-PCR验证在Huh7肝癌细胞和Huh7肝癌干细胞及10对临床肝癌组织和癌旁组织中表达差异最明显的6个miRNAs,筛选出差异具有统计学意义的miRNAs作为后续血清检测的候选miRNAs。检测和统计分析候选miRNAs在肝细胞癌患者和健康人群血清中的含量差异,分析其含量与患者部分临床病理学特征的相关性及其作为肝细胞癌诊断血清标志物的诊断效能,并分析其与肝细胞癌患者生存状况的关系。结果 (1)miR-187-3p在81例肝细胞癌患者血清中的平均相对含量为3.38±0.44,在81例健康人群血清中的平均相对含量为1.08±0.067,差异有统计学意义,χ~2=7.382,P<0.001。肝细胞癌患者血清中miR-187-3p含量和患者年龄、肿瘤大小、Child分级、AFP、ALT、门静脉癌栓和肿瘤转移情况无明显相关性,但与巴塞罗那分期相关,χ~2=4.007,P=0.049。(2)miR-187-3p诊断肝细胞癌的ROC曲线下面积为0.84,最佳灵敏度和特异度分别为76.53%和82.71%,95%CI为0.77~0.89,P=0.001。(3)42例miR-187-3p低含量组肝细胞癌患者中位无病进展期(median progression free survival,mPFS)为9.4个月,39例miR-187-3p高含量组肝细胞癌患者mPFS为2.2个月,差异有统计学意义,χ~2=23.192,P=0.001。40例miR-187-3p低含量组肝细胞癌患者中位总生存期(median overall survival,mOS)为15.6个月,39例miR-187-3p高含量组肝细胞癌患者mOS为3.8个月,差异有统计学意义,χ~2=39.074,P=0.001。结论血清miR-187-3p可能成为肝细胞癌诊断及预后判断较为特异和敏感的生物标志物。

OBJECTIVE Serum miRNA is a novel biomarker that can be used diagnosis and prognosis of cancer be- cause of its favorable specificity and stability. Various miRNAs have been used for diagnostic and prognosis of hepatocellu- lar carcinoma in recent years. In this study,we aimed to investigate the the relative expression levels of miR-187-3p which were differentially expressed between Huh7 liver cancer cell and Huh7 liver cancer stemqike cells by miRNA microarray previously, the relation between miR-187-3p expression and clinical pathological features, diagnosis of hepatocellular card noma（HCC） ,and prognosis of HCC. METHODS Serum samples of 81 HCC patients（without any treatment） and 81 healthy were collected from January 1,2012 to November 30,2016 and they were consistent in age and gender. We selected the most differentially expressed 6 miRNAs between Huh7 liver cancer stem like cell and Huh7 liver cancer cell by miR- NA microarray analysis. We were further verified these miRNAs in Huh7 liver cancer stem like cell and Huh7 liver cancer cell and 10 pairs clinical HCC specimens and adjacent non-tumorous liver tissues by qRT-PCR. Choose candidate miRNAs for next serum detection. We further detected and analyzed serum quantities of miRNAs in HCC patients and healthy sub- jects. ROC curve and AUC were used to validate the diagnostic value. RESULTS （1） The relative expression of miR-187-3p in HCC（3.38±0.44） was significantly higher than that in healthy（1.08± 0. 067,x2=7. 382, P〈0. 001）. Moreover,the level of miR-187-3p was significantly correlated with Barcelona Clinic Liver Cancer（x2 =4. 007 ,P=0. 049）, hut was no correlated with tumor size, Child-Pugh grade, AFP, ALT, portal vein tumor thromhus and metastasis. （2）ROC curve analysis showed miR-187 3p served as a potential biomarkers for HCC with an area under the ROC curve（AUC） of 0.84（95 %CI：0.77-0.89,P=0. 001,sensitivity= 76.53% ,specificity= 82.71%）. （3） HCC patients of high miR-187 3p expression level had distinctly shorter progression-free survival（median： 2.2 mouths） and a shorter overall survival（medi- an： 3.8 mouths） than that of the patients of low miR 187-3p expression level（mPFS： 9.4 mouths;mOS： 15.6 mouths）. The differences were statistically significant （mPFS： x2 = 23. 192, P = 0. 001; mOS：x2 = 39. 074, P = 0. 001）. CONCLUSION These findings indicated that the serum miR 187-3p could be used as a promising novel biomarker for the diagnosis and prognosis of HCC.